CHAPTER III

SPECIFIC ISSUES IN STS

LESSON 4: GENE THERAPY
• Human gene therapy seeks to modify or manipulate
the expression of a gene or to alter the biological
properties of living cells for therapeutic use
• Genes contain your DNA — the code that controls
much of your body's form and function, from making
you grow taller to regulating your body systems.
Genes that don't work properly can cause disease.
 GENE THERAPY

Gene therapy is a technique that modifies a person’s genes to treat or cure

disease. Gene therapies can work by several mechanisms:
• Replacing a disease-causing gene with a healthy copy of the gene
• Inactivating a disease-causing gene that is not functioning properly
• Introducing a new or modified gene into the body to help treat a
disease
 RISKS
Gene therapy has some potential risks. A gene can't easily be inserted directly into your cells.
Rather, it usually has to be delivered using a carrier, called a vector. The most common gene
therapy vectors are viruses because they can recognize certain cells and carry genetic material into
the cells' genes. Researchers remove the original disease-causing genes from the viruses, replacing
them with the genes needed to stop disease. This technique presents the following risks:

❑ Unwanted immune system reaction.Your body's immune system may see the newly
introduced viruses as intruders and attack them. This may cause inflammation and, in
severe cases, organ failure.
❑ Targeting the wrong cells. Because viruses can affect more than one type of cells, it's
possible that the altered viruses may infect additional cells — not just the targeted cells
containing mutated genes. If this happens, healthy cells may be damaged, causing other
illness or diseases, such as cancer.
❑ Infection caused by the virus. It's possible that once introduced into the body, the viruses
may recover their original ability to cause disease.
❑ Possibility of causing a tumor. If the new genes get inserted in the wrong spot in your DNA,
there is a chance that the insertion might lead to tumor formation.
• The gene therapy clinical trials underway in the U.S. are
closely monitored by the Food and Drug Administration and
the National Institutes of Health to ensure that patient safety
issues are a top priority during research.
• Currently, the only way for you to receive gene therapy is to
participate in a clinical trial. Clinical trials are research studies
that help doctors determine whether a gene therapy approach
is safe for people. They also help doctors understand the
effects of gene therapy on the body.
 SPECIFIC PROCEDURE

Your specific procedure will depend on the disease you have and the type of gene therapy
being used. For example, in one type of gene therapy:
• You may have blood drawn or you may need bone marrow removed from your
hipbone with a large needle.
• Then, in a lab, cells from the blood or bone marrow are exposed to a virus or another
type of vector that contains the desired genetic material.
• Once the vector has entered the cells in the lab, those cells are injected back into your
body into a vein or into tissue, where your cells take up the vector along with the
altered genes.
 OTHER VECTORS

Viruses aren't the only vectors that can be used to carry

altered genes into your body's cells. Other vectors being
studied in clinical trials include:
• Stem cells
• Liposomes
 STEM CELS

• Sometimes called the body’s “master cells,” stem cells are the cells that develop into blood, brain,
bones, and all of the body’s organs. They have the potential to repair, restore, replace, and
regenerate cells, and could possibly be used to treat many medical conditions and diseases.

• All medical treatments have benefits and risks. But unproven stem cell therapies can be particularly
unsafe.

• For instance, attendees at a 2016 FDA public workshop discussed several cases of severe adverse
events. One patient became blind due to an injection of stem cells into the eye. Another patient
received a spinal cord injection that caused the growth of a spinal tumor.
TYPE OS STEM CELLS
 WHERE DO ADULT STEM CELLS COME
FROM?

• Day-to-day living means the body is constantly renewing its tissues. In some parts of
the body, such as the gut and bone marrow, stem cells regularly divide to produce
new body tissues for maintenance and repair.
 WHERE DO EMBRYONIC STEM CELLS
COME FROM?
• From the very earliest stage of pregnancy, after the sperm fertilizes the
egg, an embryo forms.

• Around 3–5 days after a sperm fertilizes an egg, the embryo takes the
form of a blastocyst or ball of cells.

• The blastocyst contains stem cells and will later implant in the womb.
Embryonic stem cells come from a blastocyst that is 4–5 days old.
 WHERE DO EMBRYONIC STEM CELLS
COME FROM?
• When scientists take stem cells from embryos, these are usually extra embryos that result from in vitro
fertilization (IVF).

• In IVF clinics, the doctors fertilize several eggs in a test tube, to ensure that at least one survives. They will
then implant a limited number of eggs to start a pregnancy.
• When a sperm fertilizes an egg, these cells combine to form a single cell called a zygote.

• This single-celled zygote then starts to divide, forming 2, 4, 8, 16 cells, and so on. Now it is an embryo.

• Soon, and before the embryo implants in the uterus, this mass of around 150–200 cells is the blastocyst.
 WHERE DO EMBRYONIC STEM CELLS
COME FROM?
• The inner cell mass is where embryonic stem cells are found. Scientists call these
totipotent cells. The term totipotent refer to the fact that they have total potential to
develop into any cell in the body.

• With the right stimulation, the cells can become blood cells, skin cells, and all the other
cell types that a body needs.

• In early pregnancy, the blastocyst stage continues for about 5 days before the embryo
implants in the uterus, or womb. At this stage, stem cells begin to differentiate.
• Embryonic stem cells can differentiate into more cell types than adult stem cells.
 LIPOSOMES

• Liposomes are small artificial vesicles of spherical shape

that can be created from cholesterol and natural non-
toxic phospholipids. Due to their size and hydrophobic
and hydrophilic character(besides biocompatibility),
liposomes are promising systems for drug delivery
 RESULTS

The possibilities of gene therapy hold much promise. Clinical trials of gene therapy in
people have shown some success in treating certain diseases, such as:
• Severe combined immune deficiency
• Hemophilia – a bleeding disorder in which blood does not clot properly
• Blindness caused by retinitis pigmentosa – rare eye disease causing vision loss
• Leukemia
• But several significant barriers stand in the way
of gene therapy becoming a reliable form of
treatment, including:
• Finding a reliable way to get genetic material
into cells
• Targeting the correct cells
• Reducing the risk of side effects
• Gene therapy continues to be a very important
and active area of research aimed at developing
new, effective treatments for a variety of
diseases.
 BACTERIAL ‘NANOSYRINGE’ COULD DELIVER
GENE THERAPY TO HUMAN CELLS

• Inside the gut of a caterpillar lives a worm, and inside

the worm lurks a bioluminescent bacterium
named Photorhabdus asymbiotica, which makes the
caterpillar glow in the dark. But this nesting-doll-like
setup has another, more harmful effect: the bacteria
secrete a deadly molecular syringe, 100 nanometers
long, that latches onto the insect’s cells. Once attached
to a cell, the syringe pushes a molecular spear through
the cell’s membrane that releases a toxic payload. As its
insect host dies and decomposes, the bacteria escape to
colonize their next victim.
BACTERIAL NANOSYRINGE
(GREEN) BINDS TO INSECT
CELLS (BLUE) PRIOR TO
INJECTION OF PAYLOAD
PROTEINS.

CREDIT: JOSEPH
KREITZ/BROAD INSTITUTE OF
MIT AND HARVARD/MIT
MCGOVERN INSTITUTE FOR
BRAIN RESEARCH
 PHOTORHABDUS’S SYRINGE

• In a paper published today in Nature, researchers report refashioning Photorhabdus’s syringe—called a contractile
injection system—so that it can attach to human cells and inject large proteins into them. The work could provide a
way to deliver various therapeutic proteins into any type of cell, including proteins that can “edit” the cell’s DNA.
“It’s a very interesting approach,” says Mark Kay, a gene therapy researcher at Stanford University who was not
involved in the study. “Where I think it could be very useful is when you want to express proteins that can do
genome editing” to correct or knock out a gene that is mutated in a genetic disorder, he says.
• The nano injector could provide a critical tool for scientists interested in tweaking genes. “Delivery is probably the
biggest unsolved problem for gene editing,” says study investigator Feng Zhang, a molecular biologist at the
McGovern Institute for Brain Research at the Massachusetts Institute of Technology and the Broad Institute of
M.I.T. and Harvard. Zhang is known for his work developing the gene editing system CRISPR-Cas9. Existing
technology can insert the editing machinery “into a few tissues, blood and liver and the eye, but we
don’t have a good way to get to anywhere else,” such as the brain, heart, lung or kidney, Zhang says.
The syringe technology also holds promise for treating cancer because it can be engineered to attach to receptors on
certain cancer cells.
• It is still very early days for the technology, however. Zhang plans
to build on its efficiency as a delivery device as well as to
experiment with nonprotein payloads such as DNA and RNA.
Down the road, it will be important to test the technology in
“higher mammals,” Kay says. “There are a lot of things that work
well in mice or smaller mammals that don’t end up working as
well in nonhuman primates or humans,” he adds. And because
the injection systems consist of bacterial proteins, they could also
lead to immune reactions in humans. “We need to know: How
immunogenic is it if we put it into humans?” Zhang says. (study
investigator Feng Zhang, a molecular biologist at the McGovern
Institute for Brain Research at the Massachusetts Institute of
Technology and the Broad Institute of M.I.T. and Harvard)
 REFERENCES
• https://www.fda.gov/vaccines-blood-biologics/cellular-gene-therapy-products/what-gene-
therapy#:~:text=Gene%20therapy%20is%20a%20technique,healthy%20copy%20of%20the%20gene

• https://www.mayoclinic.org/tests-procedures/gene-therapy/about/pac-20384619

• https://www.scientificamerican.com/article/bacterial-nanosyringe-could-deliver-gene-therapy-to-
human-cells/

• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3599573/#:~:text=Liposomes%20are%20small%20
artificial%20vesicles,promising%20systems%20for%20drug%20delivery.

• https://www.fda.gov/consumers/consumer-updates/fda-warns-about-stem-cell-
therapies#:~:text=Sometimes%20called%20the%20body's%20%E2%80%9Cmaster,many%20medic
al%20conditions%20and%20diseases.

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• https://www.medicalnewstoday.com/articles/323343#sources